Rolling circle amplification and multiplex allele-specific PCR for rapid detection of katG and inhA gene mutations in Mycobacterium tuberculosis

Int J Med Microbiol. 2009 Dec;299(8):574-81. doi: 10.1016/j.ijmm.2009.05.006. Epub 2009 Jul 14.

Abstract

The aim of the study was to compare a novel, rolling circle amplification (RCA) assay for detection of common isoniazid (INH) resistance mutations in Mycobacterium tuberculosis with a multiplex allele-specific PCR (MAS-PCR) and sequencing of katG and the fabG1-inhA promoter region. One or more mutations were identified by RCA, MAS-PCR, and sequencing in 21 (68%), 22 (71%), and 23 (74%), respectively, of 31 epidemiologically unrelated INH-resistant isolates, and in none of 8 INH-susceptible isolates. The RCA assay is a rapid, inexpensive, and practical screening method for INH resistance in M. tuberculosis in countries with high prevalence of INH resistance.

Publication types

  • Evaluation Study

MeSH terms

  • Alleles
  • Bacterial Proteins / genetics*
  • Catalase / genetics*
  • Drug Resistance, Bacterial*
  • Humans
  • Microbial Sensitivity Tests / economics
  • Microbial Sensitivity Tests / methods*
  • Mutation, Missense*
  • Mycobacterium tuberculosis / drug effects*
  • Mycobacterium tuberculosis / genetics
  • Mycobacterium tuberculosis / isolation & purification
  • Nucleic Acid Amplification Techniques*
  • Oxidoreductases / genetics*
  • Polymerase Chain Reaction / economics
  • Polymerase Chain Reaction / methods*
  • Promoter Regions, Genetic
  • Sequence Analysis, DNA
  • Time Factors
  • Tuberculosis / microbiology

Substances

  • Bacterial Proteins
  • Oxidoreductases
  • Catalase
  • katG protein, Mycobacterium tuberculosis
  • InhA protein, Mycobacterium